Journal: Aging (Albany NY)
Article Title: Resemblance and differences in dietary restriction nephroprotective mechanisms in young and old rats
doi: 10.18632/aging.103960
Figure Lengend Snippet: Evaluation of the mitochondrial potential, anti-apoptotic and pro-apoptotic proteins, the intensity of mitochondrial biogenesis, and the levels of mitochondrial deacetylase SIRT-3. ( A ) Confocal microscopy of vital kidney slices loaded with mitochondrial probe TMRE. Scale bar, 100 μm; ( B ) Mitochondrial membrane potential estimated by TMRE fluorescence intensity normalized to MitoTracker Green fluorescence intensity; ( C , D ) Levels of Bcl-X L in kidney tissue of young and old rats after DR; ( E , F ) Levels of Bcl-X L in kidneys of young and old rats in response to I/R; ( G ) Comparison of Bcl-XL levels in kidney tissue of young and old rats; ( H , I ) Levels of Bcl-X s in kidney tissue of young and old rats after the DR; ( J , K ) Levels of Bcl-X s in kidney tissue of young and old rats in response to I/R; ( L ) Comparison of Bcl-XS levels in kidney tissue of young and old rats; ( M , N ) Levels of SIRT-3 in kidney tissue of young and old rats after the DR; ( O , P ) Levels of PGC-1α in kidney tissue of young and old rats after the DR. *p < 0.05 compared to AL-group. For young AL rats n=5, for young DR rats n=6, for old AL rats n=6, for old DR rats n=5. AL, ad libitum, DR, dietary restriction, IR, ischemia/reperfusion.
Article Snippet: Membranes were blocked with 5% blocking agent (Amersham Pharmacia Biotech, RPN2125V, UK) in TBS containing 0.05% Tween-20 and subsequently incubated with primary antibodies: anti-α-smooth muscle actin rabbit 1:1000 (Abcam, UK), anti-PCNA 1:1000 rabbit (Cell Signaling, USA), anti-LC3 A/B rabbit 1:1000 (Cell Signaling, USA), anti-LAMP-1 rabbit 1:1000 (Abcam, UK), anti-p62/SQSTM1 rabbit 1:1000 (Cell Signaling, USA), anti-Bcl-X rabbit 1:1000 (Cell Signaling, USA), anti-SIRT-3 rabbit 1:1000 (Cell Signaling, USA), anti-PGC-1α rabbit 1:1000 (Invitrogen, Thermo Fisher Scientific, USA), anti-β-actin mouse 1:2000 (Sigma, USA).
Techniques: Histone Deacetylase Assay, Confocal Microscopy, Fluorescence